WAe009-A-14

Cell Line

hPSCreg Name

WAe009-A-14

Cell line type

Human embryonic stem cell (hESC)

Last update

5th December 2018

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Written by
Zhiyu Zeng on 2018-12-05 03:06:40

We have generated a homozygous ATP2A2 knockout human embryonic stem cell line H9-ATP2A2-/- .H9 was transfected with Cas9 vector（Px459，Addgene,62988） which expresses a specific sgRNA sequence targeting for the exon1 of ATP2A2. After electroporation, cells were seeded on a 6-well, Matrigel precoated overnight plate and treated with 0.3ug/mL puromycin in E8 medium. Anti-puromycin clones were emerged after 4 days of electroporation. 15 clones were picked and maintained in E8 medium. DNA was extracted for genotype confirming using PCR sequencing. Clone number 4 was confirmed by homozygous deleting 1 “A” base pair. For further characterization, this clone was expanded.
H9-ATP2A2-/- grows as a clone size and expresses stem cell pluripotency hallmarks like OCT-4, SSEA4, processing a normal karyotype and the ability to differentiate into all three germ layers in vitro.

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